Our UltraScript® Reverse Transcriptase and cDNA Synthesis Kits are packed with features to provide the end-user with as many benefits as possible, simplifying their research and minimising the potential for handling errors in experiments. Read on to see how each feature can benefit your research workload.
Unbiased representation of 5′ and 3′ mRNA transcript ends
Ensures complete and accurate cDNA synthesis from full-length RNA transcripts, capturing both the 5′ and 3′ ends without bias. This enables researchers to profile gene expression comprehensively and confidently, particularly for applications like transcriptome analysis or RNA-seq.
Unbiased representation of transcript ends with UltraScript® cDNA Synthesis Kit: The UltraScript® cDNA Synthesis Kit was used to synthesise cDNA from mouse liver total RNA. Two primer pairs designed against the 5’ (purple) and 3’ (grey) ends of the (a) mouse CANX transcript (4.2 kb) and (b) the RNS18 gene (1.8 kb) were used in qPCRs with qPCRBIO SyGreen® Mix. A no RT control (blue) is shown as a negative control for each primer set.
Primer pairs up to 4 kb apart did not show any reverse transcription bias, as indicated by the overlapping amplification traces.
Reduced RNase H activity
Minimises degradation of RNA during first-strand synthesis, preserving template integrity throughout the reaction. This results in longer transcripts, higher quality cDNA and improved reproducibility, especially when working with limited or degraded RNA samples.
Thermostable reverse transcriptase 45 °C to 55 °C
Performs reliably at elevated temperatures, helping to denature RNA secondary structures that can hinder cDNA synthesis. This enhances reverse transcription efficiency and fidelity, particularly for complex or GC-rich RNA templates enabling accurate quantification of different template types.
The thermostability of UltraScript® Reverse Transcriptase was measured using the Sypro Orange fluorescence assay. The protein is incubated with Sypro Orange dye and the temperature gradually increased. The fluorescence intensity increases as the protein unfolds and the melting point is the temperature where 50% of the protein is unfolded. The DSF curve shows UltraScript Reverse Transcriptase (purple) and wildtype MMLV RT (black) at 0.1mg/ml. This experiment shows that UltraScript® Reverse Transcriptase unfolds at 54.9±0.3°C, which is 9.3°C higher than the wildtype enzyme, indicating it is more thermostable and more likely to remain active during the reaction.
As a result, where RNA regions of interest contain high secondary structure (>65% GC), incubation temperatures of up to 55°C can be used.
Advanced RNase inhibitor
Protects RNA from enzymatic degradation during the reverse transcription process, improving cDNA yield and quality. This is particularly valuable when working with precious or low-abundance RNA samples. Additionally, the inhibitor is premixed in all product formulations, thereby reducing handling time and the potential for pipetting errors.
High cDNA yields from as little as 4 pg total RNA
Delivers robust performance even with ultra-low input RNA, making it ideal for single-cell studies, rare sample types, or degraded clinical specimens. Users can achieve reliable gene expression data from minimal starting material.
Accurate reverse transcription of GC-rich templates
Effectively synthesises cDNA from RNA regions with high GC content, which are typically challenging due to stable secondary structures. This ensures complete transcript representation and avoids dropout of important gene regions.
High thermostability of UltraScript® Reverse Transcriptase enables improved cDNA yield from high GC targets.
UltraScript® cDNA Synthesis Kit was used to synthesise cDNA from mouse liver total RNA at 42°C (purple) and 55°C (gray). Primer pairs were designed against GJB2 generating an 84% GC amplicon (a) and 55% GC (b) qPCRBIO SyGreen® Mix was used for analysis. No RT reactions were used as negative controls for each primer pair (blue)
The higher temperature incubation generated more GC rich cDNA than the low cDNA amplicon
Sensitive detection of low copy number transcripts
Enables reliable reverse transcription of transcripts expressed at very low levels, supporting applications such as early disease marker detection, rare transcript quantification, and single-cell analysis.
Advanced buffer chemistry including Mg and dNTPs
Optimised reaction conditions with pre-balanced magnesium and nucleotide concentrations ensure efficient enzyme performance and consistent results. This simplifies setup and reduces variability between reactions.
Both premixed or separate oligo cDNA synthesis kits available
Provides flexible formats to suit different experimental needs. Users can choose a convenient all-in-one mix for routine workflows or opt for separate components to customise reactions for complex or specialised applications.
Fast cDNA synthesis in 15 to 30 mins
Speeds up the reverse transcription workflow, enabling users to go from RNA to cDNA in as little as 15 minutes. This saves valuable time in high-throughput or time-sensitive experiments without compromising on cDNA quality.
